Illuminating genetic interactions that affect lipid content in adipocytes

项目摘要 肥胖和相关的代谢紊乱是影响全球数百万人的重大医疗危机 而且没有长期的治疗方法。我们目前关于细胞如何调节新陈代谢的大部分知识 是通过单基因驱动假说获得的。然而，像许多其他疾病一样，肥胖也是由多基因决定的。 紊乱，其中多个基因相互作用，导致表型，这使得不可能建立 致病等位基因。如果我们要在治疗肥胖症和代谢性疾病方面取得任何进展 世纪，我们需要大胆、有创意的想法来处理细胞途径，而不是单个基因 尖端高吞吐量方法的优势。 截至2016年，全球估计有超过19亿成年人超重，6.5亿成年人肥胖。 肥胖的经典特征是脂肪组织的病理发展和分化，这是一种 人体内关键的储能器官。脂肪组织在脂滴中储存能量--直到最近才被发现 具有复杂和动态功能的细胞器。脂滴可以在大小上发生巨大变化 对卡路里摄入或新陈代谢变化的反应。由于目前FDA批准的减肥药作用于 食欲或代谢调节而不是直接对脂肪组织脂类进行调节，我们需要确定途径 负责脂滴的生物生成和分解，以改进现有的治疗方法。令人惊讶的是，几乎没有 已知影响人类脂滴的途径。 我将利用CRISPR系统建立一个新型的高通量、高内容的差异化筛选平台 脂肪细胞。由于大多数脂质修饰基因都是在非哺乳动物模型系统中发现的，例如 线虫、果蝇和酿酒酵母利用siRNA方法，我的目标是开发一种新的优越的CRISPR 哺乳动物系统中的筛选平台。我将利用我们实验室拥有的可诱导CRISPRi小鼠 开发用于在原代脂肪细胞中进行单基因和双基因扰动筛选。因此，我提出的 研究策略将不仅阐明新的基因，而且重要的是阐明基因的相互作用和途径 负责脂滴的形成和分解，为减肥治疗提供新的靶点。

Project Abstract Obesity and associated metabolic disorders are a major health care crisis that affect millions of people worldwide and have no long-term treatment. Most of our current knowledge on how cells regulate their metabolism has been obtained using single gene-driven hypothesis. However, obesity like many other diseases is a polygenic disorder, in which multiple genes interact, contributing to phenotype, which makes it impossible to establish the causative allele. If we are to make any headway towards curing obesity and metabolic diseases during this century, we need bold, creative ideas to tackle cellular pathways as oppose to individual genes that take advantage of cutting-edge high-throughput approaches. As of 2016, over 1.9 billion adults worldwide were estimated to be overweight and 650 million adults to be obese. Obesity is classically characterized by pathological development and differentiation of adipose tissue, which is a key energy storage organ in the body. Adipose tissue stores energy in lipid droplets- until recently unappreciated cellular organelles with complex and dynamic functions. Lipid droplets can drastically change in size as a response to changed calorie intake or metabolism. Since currently FDA-approved anti-obesity drugs act on appetite or metabolism regulation and not on adipose tissue lipids directly, we need to identify pathways responsible for lipid droplet biogenesis and breakdown to improve current therapeutics. Surprisingly, very little is known about the pathways that affect lipid droplets in humans. I will use a CRISPR system to establish a novel high-throughput high-content screening platform in differentiated adipocytes. Since most lipid modifying genes have been discovered in non-mammalian model systems such as C. elegans, Drosophila and S. cerevisiae using siRNA approaches, I aim to develop a novel superior CRISPR screening platform in mammalian system. I will take advantage of inducible CRISPRi mouse our lab has developed to perform a single and dual genetic perturbation screen in primary adipocytes. Thus, my proposed research strategy will illuminate not only new genes but importantly genetic interactions and pathways responsible for formation and breakdown of lipid droplets and provide new targets for anti-obesity therapy.